Project Summary/Abstract (Overall) Exposures to environmentally relevant chemicals, drugs, medications, and the ingestion of nutrients, and how they perturb endogenous metabolism have been linked with many types of adverse health outcomes. Our Hub has the resources, skilled staff, and knowledge to immediately provide Exposome services to clients of the Human Health Exposure Analysis Resource (HHEAR) Program (RFA-ES-18-012). We initiated our Untargeted Analysis Core in 2012 through a grant from the NIH Common Fund (C-F) Metabolomics Program, and have established and applied metabolomics methods to hundreds of studies with over 30 organizations across the United States. These studies have included analysis of a wide range of biospecimens (e.g., urine, serum, plasma, sweat, feces) with anchors to numerous types of health outcomes in studies of maternal and child health, kidney disease, cardiovascular disease, obesity and diabetes, and response to treatment. The success of the NIH C-F Eastern Regional Comprehensive Metabolomics Resource Core (ERCMRC) led to our establishing a Children's Health Exposure Analysis Resource (CHEAR) Hub as part of the NIEHS CHEAR Program. Under the CHEAR Program, we successfully executed our goals, which included a) expanding our services to include high-throughput analysis of the Exposome using high-resolution Orbitrap mass spectrometry, b) establishing an extensive in-house physical standards library of exogenous and endogenous compounds run on the untargeted platform, and c) participating in cross laboratory comparisons to reveal the range of environmentally relevant analytes identified on our untargeted platform. Our untargeted methods result in tens of thousands of signals that are annotated by matching to public databases, and annotated and identified against our in-house physical standards library, using Big Data Analytics. Our core led the development of an evidence-based Ontology System for the CHEAR Program, which ensures that the confidence in annotations and assignments will be clearly communicated between Hubs. We have in place over 100 fine-tuned protocols that span the workflow from receipt of samples through reporting data, and that have quality control/assurance procedures in place for all steps of the workflow. The HHEAR Untargeted Hub will work within the HHEAR Network and other HHEAR Hubs, the HHEAR Data Analysis Center (DAC), and the HHEAR Coordinating Center (CC) to provide client services for Untargeted Analysis, and to work across the consortium to harmonize and integrate results. The HHEAR Hub will be led under a Multiple PI Leadership Plan by Dr. Timothy Fennell (Director, Administrative Core and the Development Core), Dr. Susan Sumner (Director, Untargeted Analysis Resource), and Dr. Xiuxia Du (leader, Computational Exposome workflow). We are ready to provide untargeted analysis for epidemiology investigations using a wide range of biospecimens and state-of-the-art technologies, and analysis approaches to reveal the influence of the environment on disease.